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The spleens of mice that demonstrated the highest antibody titres as determined by ELISA using the peptide immunogen were fused with myeloma cells.
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Nine weeks after the initial booster, rabbits were bled and serum was tested by ELISA using the peptide as an immunogen.
EAE experiments were performed as described (Stromnes & Goverman, 2006) using the peptide MOG35 55 as an immunogen.
The experiments presented in Fig. 1 conclusively demonstrate that activation of the MAPKs ERK, JNK and p38 can be detected in human B cells in a specific manner that is blocked with the peptide immunogen used to make the phosphospecific antibodies.
(A) Alignment of the peptide immunogen (from human Smad2) used to generate the anti-Smad2 antibody with zebrafish Smad2 protein sequence.
Specificity of the NDRG1 antibody from Zymed® was confirmed by blocking with the peptide immunogen.
A mono-specific PA28γ antibody recognizing C- and N-terminal regions of the peptide immunogen was purified from polyclonal rabbit serum via a PA28γ peptide column.
The antibodies were purified from sheep serum by affinity chromatography on CH-Sepharose to which the peptide immunogen had been covalently coupled.
Indeed, all reported antibodies directed at phosphorylated TDP-43 within this search are generated using a peptide immunogen.
The peptide immunogens were coated onto microtiter plates.
To detect these products, polyclonal antibodies were raised in rabbits using a synthetic peptide immunogen corresponding to the C-terminal ten amino acids of the mutant tag with an N-terminal Cys residue to facilitate chemical crosslinking (NH3-CSSQQAFALDD-COOH).
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